Liu Bo-Tong, Li Quan, Sun Kai, Pan Chun-Shui, Huo Xin-Mei, Huang Ping, Yan Li, He Qi-Hua, Zhong Li-Jun, Wang Yuan, Hu Meng-Lei, Li An-Qing, Jiao Ying-Qian, Zhang Shuang, Wang Xiao-Yi, Liu Jian, Han Jing-Yan
Department of Integration of Chinese and Western Medicine, School of Basic Medical Sciences, Peking University, Beijing, China.
Tasly Microcirculation Research Center, Peking University Health Science Center, Beijing, China.
Front Pharmacol. 2024 Aug 1;15:1421635. doi: 10.3389/fphar.2024.1421635. eCollection 2024.
Angong Niuhuang Wan (AGNHW), developed during the Qing dynasty (18th century) for the treatment of consciousness disturbances caused by severe infections, has been used to treat brain edema caused by ischemia‒reperfusion. However, it remains unclear whether AGNHW can ameliorate vascular-origin brain edema caused by lipopolysaccharides (LPS). This study explored the ameliorative effects of AGNHW on LPS-induced cerebrovascular edema in mice, as well as the potential underlying mechanisms.
A cerebrovascular edema model was established in male C57BL/6N mice by two intraperitoneal injections of LPS (15 mg/kg), at 0 and 24 h. AGNHW was administered by gavage at doses of 0.2275 g/kg, 0.455 g/kg, and 0.91 g/kg, 2 h after LPS administration. In control mice, normal saline (NS) or AGNHW (0.455 g/kg) was administered by gavage 2 h after intraperitoneal injection of NS. The survival rate, cerebral water content, cerebral venous FITC-dextran leakage, Evans blue extravasation, and expression of vascular endothelial cadherin (VE-cadherin), zonula occludens-1 (ZO-1), claudin-5, phosphorylated caveolin-1 (CAV-1), and cytomembrane and cytoplasmic aquaporin 1 (AQP1) and aquaporin 4 (AQP4) were evaluated. The cerebral tissue phosphoproteome, blood levels of AGNHW metabolites, and the relationships between these blood metabolites and differentially phosphorylated proteins were analyzed.
AGNHW inhibited the LPS-induced decrease in survival rate, increase in cerebral water content, decrease in VE-Cadherin expression and increase in phosphorylated CAV-1 (-CAV-1). AGNHW treatment increased the expression of AQP4 on astrocyte membrane after LPS injection. AGNHW also inhibited the LPS-induced increases in the phosphorylation of 21 proteins, including protein kinase C-α (PKC-α) and mitogen-activated protein kinase 1 (MAPK1), in the cerebral tissue. Eleven AGNHW metabolites were detected in the blood. These metabolites might exert therapeutic effects by regulating PKC-α and MAPK1.
AGNHW can ameliorate cerebrovascular edema caused by LPS. This effect is associated with the inhibition of VE-Cadherin reduction and CAV-1 phosphorylation, as well as the upregulation of AQP4 expression on the astrocyte membrane, following LPS injection.
安宫牛黄丸（AGNHW）于清朝（18世纪）研发，用于治疗严重感染引起的意识障碍，现也被用于治疗缺血再灌注引起的脑水肿。然而，AGNHW是否能改善脂多糖（LPS）引起的血管源性脑水肿仍不清楚。本研究探讨了AGNHW对LPS诱导的小鼠脑血管水肿的改善作用及其潜在机制。
通过在0小时和24小时腹腔注射两次LPS（15mg/kg），在雄性C57BL/6N小鼠中建立脑血管水肿模型。在LPS给药后2小时，通过灌胃给予AGNHW，剂量分别为0.2275g/kg、0.455g/kg和0.91g/kg。在对照小鼠中，腹腔注射生理盐水（NS）后2小时，通过灌胃给予生理盐水或AGNHW（0.455g/kg）。评估存活率、脑含水量、脑静脉FITC-葡聚糖渗漏、伊文思蓝外渗以及血管内皮钙黏蛋白（VE-钙黏蛋白）、紧密连接蛋白1（ZO-1）、闭合蛋白5、磷酸化小窝蛋白1（CAV-1）以及细胞膜和细胞质水通道蛋白1（AQP1）和水通道蛋白4（AQP4）的表达。分析脑组织磷酸化蛋白质组、AGNHW代谢产物的血药浓度以及这些血药代谢产物与差异磷酸化蛋白之间的关系。
AGNHW抑制了LPS诱导的存活率降低、脑含水量增加、VE-钙黏蛋白表达降低以及磷酸化CAV-1（-CAV-1）增加。AGNHW治疗增加了LPS注射后星形胶质细胞膜上AQP4的表达。AGNHW还抑制了LPS诱导的脑组织中包括蛋白激酶C-α（PKC-α）和丝裂原活化蛋白激酶1（MAPK1）在内的21种蛋白的磷酸化增加。在血液中检测到11种AGNHW代谢产物。这些代谢产物可能通过调节PKC-α和MAPK1发挥治疗作用。
AGNHW可以改善LPS引起的脑血管水肿。这种作用与抑制LPS注射后VE-钙黏蛋白减少和CAV-1磷酸化以及上调星形胶质细胞膜上AQP4表达有关。
